Anti-Th17 and anti-Th2 responses effects of hydro-ethanolic extracts of Aframomum melegueta, Khaya senegalensis and Xylopia aethiopica in hyperreactive onchocerciasis individuals' peripheral blood mononuclear cells.

Hyperreactive onchocerciasis (HO) is characterized by a severe skin inflammation with elevated Th17-Th2 combined responses. We previously demonstrated the anthelminthic activity of Aframomum melegueta (AM), Xylopia aethiopica (XA) and Khaya senegalensis (KS) used by traditional healers to treat helminthiasis in the endemic area of Togo. However, their effect on severe onchocerciasis is poorly investigated. The present study aimed to investigate the anti-Th17 and anti-Th2 effects of hydro-ethanolic extracts of AM, XA and KS during HO. Onchocerca volvulus-infected individuals were recruited in the Central region of Togo in 2018. Isolated peripheral blood mononuclear cells (PBMCs) from both generalized onchocerciasis (GEO) and HO forms were activated with anti-CD3 and anti-CD28 monoclonal antibodies in the presence or absence of the hydro-ethanolic extracts of AM, XA and KS as well as their delipidated, deproteinized and deglycosylated fractions. After 72 hours, cytokines were assayed from cell culture supernatants. Then, flow cytometry was used to investigate the effects of the extracts on cell activation, proliferation, intracellular cytokines and T cells transcription factors. The production of both Th17 and Th2 cytokines IL-17A and IL-5 were significantly inhibited upon T-cell receptor (TCR) activation in the presence of the hydro-ethanolic extracts of AM, XA and KS in HO individuals' PBMCs in vitro. AM and XA inhibited CD4+RORC2+IL-17A+ and CD4+GATA3+IL-4+ cell populations induction. This inhibition was not Th1 nor Treg-dependent since both IFN-γ and IL-10 were also inhibited by the extracts. AM and XA did not interfere with T cell activation and proliferation for their inhibitory pathways. Lipid and protein compounds from AM and XA were associated with the inhibition of IL-17A. This study showed that in addition to their anthelminthic effects, hydro-ethanolic extracts of Aframomum melegueta, Xylopia aethiopica and Khaya senegalensis could downregulate both Th17 and Th2 responses and prevent the severe skin disorder observed.

[Corrigendum: Ethnobotanical study of plants used in the treatment of diabetes in the traditional medicine of Maritime Region, Togo]. / Corrigendum: Étude ethnobotanique des plantes utilisées dans le traitement du diabète dans la médecine traditionnelle de la région Maritime du Togo.

[This corrects the article DOI: 10.11604/pamj.2015.20.437.5660.][This corrects the article DOI: 10.11604/pamj.2015.20.437.5660.].

Molecular detection of Mycobacterium ulcerans in the environment and its relationship with Buruli ulcer occurrence in Zio and Yoto districts of maritime region in Togo.

BACKGROUND: Buruli Ulcer (BU) is a neglected tropical skin infection caused by Mycobacterium ulcerans. Residence near aquatic areas has been identified as an important source of transmission of M. ulcerans with increased risk of contracting Buruli ulcer. However, the reservoir and the mode of transmission are not yet well known. The aim of this study was to identify the presence of M. ulcerans in the environment and its relationship with Buruli ulcer occurrence in Zio and Yoto districts of the maritime region in south Togo. METHODS: A total of 219 environmental samples including soil (n = 119), water (n = 65), biofilms/plants (n = 29) and animals' feces (n = 6) were collected in 17 villages of Zio and Yoto districts of the maritime region in Togo. DNA of M. ulcerans including IS2404 and IS2606 insertions sequences and mycolactone ketoreductase-B gene (KR-B) was detected using real time PCR amplification (qPCR) technique. In parallel, clinical samples of patients were tested to establish a comparison of the genetic profile of M. ulcerans between the two types of samples. A calibration curve was generated for IS2404 from a synthetic gene of M. ulcerans Transposase pMUM001, the plasmid of virulence. RESULTS: In the absence of inhibition of the qPCR, 6/219 (2.7%) samples were tested positive for M. ulcerans DNA containing three sequences (IS2404/IS2606/KR-B). Positive samples of M. ulcerans were consisting of biofilms/plants (3/29; 10.3%), water (1/65; 1.7%) and soil (2/119; 1.5%). Comparative analysis between DNA detected in environmental and clinical samples from BU patients showed the same genetic profile of M. ulcerans in the same environment. All these samples were collected in the environment of Haho and Zio rivers in the maritime region. CONCLUSION: This study confirms the presence of M. ulcerans in the environment of the Zio and Yoto districts of the maritime region of Togo. This may explain partially, the high rates of Buruli ulcer patients in this region. Also, water, plants and soil along the rivers could be possible reservoirs of the bacterium. Therefore, Haho and Zio rivers could be potential sources of infection with M. ulcerans in humans in these districts.

Risk factors for Mycobacterium ulcerans infection (Buruli Ulcer) in Togo ─ a case-control study in Zio and Yoto districts of the maritime region.

BACKGROUND: Buruli ulcer (BU) is a neglected mycobacterial skin infection caused by Mycobacterium ulcerans. This disease mostly affects poor rural populations, especially in areas with low hygiene standards and sanitation coverage. The objective of this study was to identify these risk factors in the districts of Zio and Yoto of the Maritime Region in Togo. METHODS: We conducted a case-control study in Zio and Yoto, two districts proved BU endemic from November 2014 to May 2015. BU cases were diagnosed according to the WHO clinical case definition at the Centre Hospitalier Régional de Tsévié (CHR Tsévié) and confirmed by Ziehl-Neelsen (ZN) microscopy and IS2404 polymerase chain reaction (PCR). For each case, up to two controls matched by sex and place of residence were recruited. Socio-demographic, environmental or behavioral data were collected and conditional logistic regression analysis was used to identify and compare risk factors between BU cases and controls. RESULTS: A total of 83 cases and 128 controls were enrolled. The median age was 15 years (range 3-65 years). Multivariate conditional logistic regression analysis after adjustment for potential confounders identified age (< 10 years (OR =11.48, 95% CI = 3.72-35.43) and 10-14 years (OR = 3.63, 95% CI = 1.22-10.83)), receiving insect bites near a river (OR = 7.8, 95% CI = 1.48-41.21) and bathing with water from open borehole (OR = 5.77, (1.11-29.27)) as independent predictors of acquiring BU infection. CONCLUSIONS: This study identified age, bathing with water from open borehole and receiving insect bites near a river as potential risk of acquiring BU infection in Zio and Yoto districts of the Maritime Region in south Togo.

THERAPEUTIC POTENTIALS OF ETHANOLIC EXTRACT OF LEAVES OF HOLARRHENA FLORIBUNDA (G. DON) DUR. AND SCHINZ (APOCYNACEAE).

BACKGROUND: Holarrhena floribunda is a plant of wide usage in the Togolese folk medicine. A previous ethnobotanical survey on the latex plants of the Maritime region of the country revealed that this plant was included in several recipes curing malaria and microbial infections. Therefore, this study aimed to seek for the effectiveness of the ethanolic extract of the plant in the treatment of these diseases. METHODS: The antimicrobial test was performed using the agar well-diffusion and the NCCLS broth microdilution methods, while the in vivo antimalarial activity was evaluated following the four-day suppressive test of Peters. The acute toxic effects of the extract were monitored after a single oral dose (5,000 mg/kg body weight) administration in NMRI mice. RESULTS: The results indicated that the ethanolic extract of leaves of H. floribunda was active on Staphylococcus aureus ATCC 29213 and clinical strains of Staphylococcus aureus, Salmonella typhi and Klebsiella pneumoniae with MICs ranging from 0.62 to 1.25 mg/mL. The extract also showed significant parasitaemia suppression in a dose-dependent manner. In the acute toxicity assay, the oral administration of the extract to the mice did not affect the relative weight of vital organs, and there were no signs of toxicity or death during the study period. The LD50 of the tested extract was found to be greater than 5,000 mg/kg, indicating its safety. CONCLUSION: This study demonstrates the antibacterial and antimalarial activities of leaves of H. floribunda and then, supports its medicinal use in the treatment of microbial infections.

Ethnobotanical Study of Medicinal Plants Used for the Treatment of Malaria in the Plateau Region, Togo.

BACKGROUND: In Togo, malaria constitutes a major public health problem but, until now, the population still mostly relies on herbal medicine for healing. This study aimed to document medicinal plants used for malaria therapy in the Plateau region of the country. METHODOLOGY: Semi-structured questionnaire interviews were used to gather ethnobotanical and sociodemographic data from traditional healers of the study area. RESULTS: A total of 61 plants species belonging to 33 families were found to be in use for malaria therapy in the Plateau region. Caesalpiniaceae were the most represented family with 7 species, followed by Euphorbiaceae and Poaceae with 4 species each. According to the relative frequency of citation (RFC), Newbouldia laevis Seem. (RFC =0.52), Sarcocephalus latifolius (Sm.) E.A. Bruce (RFC =0.48), Acanthospermum hispidum DC. (RFC =0.43), and Senna siamea (Lam.) H.S. Irwin and Barneby (RFC =0.40) were the most cited in the treatment of malaria in the traditional medicine in the Plateau region. The parts of plants used could either be the barks, roots, leaves, or whole plants. The recipes also could be a combination of various species of plants or plant parts. CONCLUSION: This study highlights the potential sources for the development of new antimalarial drugs from indigenous medicinal plants found in the Plateau region of Togo. Such results could be a starting point for in vitro antimalarial screenings. SUMMARY: 61 plants species from 33 families are use for malaria therapy in the Plateau region of TogoThe main families are Caesalpiniaceae Euphorbiaceae and PoaceaeThe most used species are Newbouldia laevis Seem. (RFC = 0.52), Sarcocephalus latifolius (Sm.) E.A. Bruce (RFC = 0.48), Acanthospermum hispidum DC. (RFC = 0.43), and Senna siamea (Lam.) H.S. Irwin and Barneby (RFC = 0.40) Abbreviations Used: RFC: Relative frequency of citation, FC: Frequency of citation, Dec: Decoction, Orl: Oral route, Mac: Maceration, Jui: Juice, Inf: Infusion, Sau: Sauce, Kne: Kneading, Le: Leaves, Rt: Roots, Wp: Whole plant, St: Stem, Stb: Stem bark, Rh: Rhizome, Fr: Fruits, Pf: Plasmodium falciparum, IC50: Concentration of extract killing 50% parasites.

Ethnobotanical Study of Latex Plants in the Maritime Region of Togo.

BACKGROUND: In Togo, a little is known about latex plants of the flora used for medicinal purposes. OBJECTIVE: The aim of this study was to identify the latex plant species and their medicinal uses in the Maritime Region of Togo. MATERIALS AND METHODS: The methodology was based on ethnobotanical semi-structural individual interviews of 220 informants. Quantitative ethnobotanical index was used to analyze the data. RESULTS: A total of 33 latex plants species were recorded, from 12 botanical families and 24 genera. The most represented families were Euphorbiaceae and Moraceae with eight species each. The relative importance (RI) value of each species and the informant consensus factor (ICF) of the ailments categories showed that Pergularia daemia (Forssk.) Chiov. (RI = 2.00) and Euphorbia hirta L. (RI = 1.91) were the most versatile in relation to their uses, and infectious diseases (ICF = 0.922) were the category with the greatest consensus among 17 categories. CONCLUSION: These latex plants of Togolese flora are variously used in traditional medicine and it would be important to undertake further investigations in phytochemistry, pharmacology, and toxicology to validate their uses. Abbreviations Used: UV: use value ICF: informant consensus factor RI: relative importance PP: pharmacological properties attributed to a species for a specific ailments AC: ailment categories treated by a given species.

An ethnobotanical study of plants used to treat liver diseases in the Maritime region of Togo.

ETHNOPHARMACOLOGICAL RELEVANCE: In Togo, many persons still rely on plants for healing, however very little is known about the medicinal practices of the indigenous people. The present study aimed to document the medicinal plant utilization for the management of liver diseases in the Maritime region of the country. METHODOLOGY: This was an ethnobotanical survey conducted in the Maritime region of Togo from June to August 2015. The data were gathered from 104 traditional healers (TH) by direct interviews using a semi-structured questionnaire. The calculated use values (UV) were used to analyze the importance of the cited plants. RESULTS: A total of 99 plant species belonging to 88 genera and 49 families were cited by the TH as curing the hepatic diseases. The most represented families were Caesalpiniaceae with 8 species, followed by Euphorbiaceae with 7 species, Apocynaceae and Asteraceae with 6 species each. The highest UV were recorded with Gomphrena celosioides (0.13), Xylopia ethiopica (0.12), Senna occidentalis (0.12), Bridelia ferruginea (0.12), Cymbopogon citratus (0.12), Kigellia Africana (0.09), Cassia sieberiana (0.08) and Sanseviera liberica (0.08), showing their importance in the management of liver dysfunction in the surveyed region. The main used parts were the leaves, followed by the roots, the whole plant, the rhizome and the bark, accounting for more than 10% each. The herbal medicines were mostly prepared in the form of decoction and administrated by oral route. CONCLUSION: This study showed that Maritime region of Togo has an important plant biodiversity that is exploited by the indigenous TH. However, some plants cited by the TH have not been studied for their possible hepatoprotective effects. These plants are therefore a starting point for biological screenings.

Medicinal plants use in central Togo (Africa) with an emphasis on the timing.

BACKGROUND: Plant-based remedies continue to play a key role in the health care of people in Togo; however, there is a lack of published data in medicinal plants and medical practices of the people in the country. OBJECTIVE: This study was aimed at documenting the plant utilization in the Tem folk medicine in the central region of Togo. MATERIALS AND METHODS: An ethnobotanical survey was conducted with traditional healers in the central region of Togo using a semi-structured questionnaire. RESULTS: This study demonstrated that local specialists in the central region of Togo tend to agree with each other in terms of the plants used to treat diabetes (ICF = 0.38), infertility, and abdominal pains (ICF = 0.33), but cite a much more diverse groups of plants to treat problems related to arterial hypertension, sickle cell disease, and abscess. They use 144 herbal concoctions made of 72 plants, distributed among 36 botanical families. The Euphorbiaceae family with eight species was best represented in terms of the number of species. The species with the highest use value were Khaya senegalensis (Desr.) A. Juss. (Meliaceae) (UV = 0.36), Anthocleista djalonensis A. Chev. (Gentianaceae) (UV = 0.27), Trichilia emetica Vahl (Meliaceae) (UV = 0.25), and Sarcocephalus latifolius (Sm.) E. A. Bruce (Rubiaceae) (UV = 0.21). They also rely on the timing in the plant processing and the administration of herbal remedies. CONCLUSION: All these findings are based on empirical observations; laboratory screenings are needed to check the effectiveness of these plants.

Screening Togolese medicinal plants for few pharmacological properties.

BACKGROUND: Terminalia macroptera Guill. et Perr. (Combretaceae), Sida alba L. (Malvaceae), Prosopis africana Guill et Perr. Taub. (Mimosaceae), Bridelia ferruginea Benth. (Euphorbiaceae), and Vetiveria nigritana Stapf. (Asteraceae) are traditionally used in Togolese folk medicine to treat several diseases including microbial infections. OBJECTIVE: This study aimed to investigate the antimicrobial, antioxidant, and hemolytic properties of the crude extracts of the above-mentioned plants. MATERIALS AND METHODS: The antimicrobial and the antioxidant activities were assayed using the NCCLS microdilution method and the DPPH free radical scavenging, respectively. Human A+ red blood cells were used to perform the hemolytic assay. Phenolics were further quantified in the extracts using spectrophotometric methods. RESULTS: Minimal inhibitory concentrations in the range of 230-1800 µg/ml were recorded in the NCCLS broth microdilution for both bacterial and fungal strains with methanol extracts. The DPPH radical scavenging assay yielded interesting antioxidant activities of the extracts of P. africana and T. macroptera (IC(50) values of 0.003 ± 0.00 µg/ml and 0.05 ± 0.03 µg/ml, respectively). These activities were positively correlated with the total phenolic contents and negatively correlated with the proanthocyanidin content of the extracts. The hemolytic assay revealed that great hemolysis occurred with the methanol extracts of T. macroptera, S. longepedunculata, and B. ferruginea. CONCLUSION: These results support in part the use of the selected plants in the treatment of microbial infections. In addition, the plant showed an interesting antioxidant activity that could be useful in the management of oxidative stress.

Ethnobotanical study of medicinal plants used in the management of diabetes mellitus and hypertension in the Central Region of Togo.

CONTEXT: The Tem tribe in the Central Region of Togo is a population with an extensive knowledge of medicinal plants. However, little is known about their medical practices, principally the use of plants in the management of diabetes mellitus (DM) and hypertension (HTN). OBJECTIVE: The present study documented the indigenous medicinal plant utilization for the management of DM and HTN in the Togo Central Region. METHODOLOGY: From March to October 2010, 55 traditional healers were interviewed about their knowledge on the use of plants for DM and HTN treatment. RESULTS: The results revealed that 35/55 (63.64%) healers had treated at least one case of DM and/or HTN. They highlighted the use of 64 species belonging to 31 families in the treatment of DM and/or HTN. The most used plants against diabetes were Psidium guajava L. (Myrtaceae), Khaya senegalensis A. Juss. (Meliaceae), Sarcocephalus latifolius (Sm.) E.A. Bruce (Rubiaceae), Annona muricata L. (Annonaceae), Bridelia ferruginea Benth. (Phyllanthaceae), and Securidaca longepedunculata Fresen. (Polygalacae), while Allium sativum L. (Liliaceae) and Parkia biglobosa Benth. (Fabaceae), followed by Khaya senegalensis A. Juss. (Meliaceae), Gardenia ternifolia Schumach. (Rubiaceae), and Persea americana Mill. (Lauraceae) were the most commonly cited as antihypertensive. CONCLUSION: The issue revealed that traditional healers of the above mentioned region have basic knowledge regarding herbal medicine for DM and HTN in comparison with previous published reports. Further pharmacological screening of the identified plants should be conducted to ascertain the effectiveness of these plants.

Antimicrobial, antiplasmodial, haemolytic and antioxidant activities of crude extracts from three selected Togolese medicinal plants.

OBJECTIVE: To investigate the antioxidant, antimicrobial, antiplasmodial, acute toxicity and haemolytic activities of methanolic extracts of three plants. Phytochemical analysis to determine the phenolic contents was also carried out. METHODS: The 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging, NCCLS broth microdilution and Plasmodium Lactate Dehydrogenase (pLDH) assays were used to determine antioxidant, antimicrobial and antiplasmodial activities, respectively. Haemolysis assay was conducted on A(+) human red blood cells and acute toxicity on male Swiss albino mice. Phenolics were quantitatively determined using spectrophotometric methods. RESULTS: The DPPH assay yielded interesting antioxidant activities of methanolic extract of Parinari curatellifolia (P. curatellifolia) and Entada africana (E. africana) (IC(50) were 0.20±0.01 µg/mL and 0.47±0.01 µg/mL, respectively). This activity was highly correlated with phenolic contents of extracts. The antimicrobial tests displayed minimal inhibitory concentrations (MICs) values ranging from 0.90 to 1.80 mg/mL for Serratia marcescens (S. marcescens) the most susceptible bacterial strain. MIC value was 1.20 mg/mL for susceptible fungal strains including Mucor rouxi (M. rouxi), Fusarium oxyporum (F. oxyporum) and Rhizopus nigricans (R. nigricans). pLDH assay showed moderate antiplasmodial activity of Balanites aegyptiaca (B. aegyptiaca) (IC(50) = 24.56±3.45 µg/mL), however this extract was highly haemolytic and toxic in mice (LD(50) = 625±128 mg/kg). CONCLUSIONS: Our results support in part the use of the selected plants in the treatment of microbial infections. In addition the plant showed interesting antioxidant activity that could be useful in the management of oxidative stress.

Sub-Saharan Rubiaceae: a review of their traditional uses, phytochemistry and biological activities.

Rubiaceae family is a large family of 630 genera and about 13000 species found worldwide, especially in tropical and warm regions. These plants are not only ornamental but they are also used in African folk medicine to treat several diseases. Based on online published data and library bibliographic research, we herein reported accumulated information related to their traditional usages in sub-Saharan traditional medicine, their chemical composition and the screened pharmacological activities. Indeed, more than 60 species are used for more than 70 medicinal indications including malaria, hepatitis, eczema, oedema, cough, hypertension, diabetes and sexual weakness. Through biological screening following leads supplied with traditional healers, many of these plants exhibited antimalarial, antimicrobial, antihypertension, antidiabetic, antioxidant and anti-inflammatory activities. Bioactive compounds including indole alkaloids, terpenoids and anthraquinones have been isolated from these bioguided fractionation studies. It is evidence that great attention has been paid to species such as Nauclea latifolia, Morinda lucida, Mitragyna inermis and Crossopteryx febrifuga; however, several compounds should be waiting to be discovered since none of these plants has been systematically investigated for its biochemical composition. According the current global health context with the recrudescence of HIV, much effort should be oriented towards this virus when screening Rubiaceae.

Herbal remedies and their adverse effects in Tem tribe traditional medicine in Togo.

In Africa, up to 80% of the population relies on herbal concoctions for their primarily health care. In Togo, western Africa, Tem tribe is a population with old knowledge of medicinal plants, however, still very little is known about their medical practices. The present study was conducted to access for the apprehension of adverse effects of traditional remedies by Tem traditional healers (TH). Enquiry was performed by interviews with healers from August to October 2007 in Tchaoudjo prefecture (Togo). The study allowed us to interview 54 TH including 41(75.93%) males and 13(24.07%) females, who cited 102 recipes assumed to have adverse effects. The recipes were used alone to cure several diseases including haemorrhoids (22.55%), female sexual disorders and infertility (21.57%), gastrointestinal disorders (18.63%), and malaria (6.86%). A total of 34 plants belonging to 21 families were cited to be components of the recipes. Euphorbiaceae and Mimosaceae families were the most represented, however, Nauclea latifolia, Khaya senegalensis, Pseudocedrela kotschyi and Xeroderris stuhlmannii were the main components of recipes linked to adverse effects. A total of 20 adverse effects were linked to the administration of theses drugs, and among them; diarrhoea, abdominal pains, polyuria, general weakness and vomiting were the most frequently encountered. These findings were in accordance with several reports of the literature concerning medicinal plants, although they were based on empirical observations. Laboratory screenings are needed to access for the effectiveness as well as the possible toxic effects of the recipes.